Carboxy-Terminal Cementum Protein 1-Derived Peptide 4 (cemp1-p4) Promotes Mineralization through wnt/β-catenin Signaling in Human Oral Mucosa Stem Cells

Abstract

Human cementum protein 1 (CEMP1) is known to induce cementoblast and osteoblast differentiation and alkaline phosphatase (ALP) activity in human periodontal ligament-derived cells in vitro and promotes bone regeneration in vivo. CEMP1′s secondary structure analysis shows that it has a random-coiled structure and is considered an Intrinsic Disordered Protein (IDP). CEMP1′s short peptide sequences mimic the biological capabilities of CEMP1. However, the role and mechanisms of CEMP1′s C-terminal-derived synthetic peptide (CEMP1-p4) in the canonical Wnt/β-catenin signaling pathway are yet to be described. Here we report that CEMP1-p4 promotes proliferation and differentiation of Human Oral Mucosa Stem Cells (HOMSCs) by activating the Wnt/β-catenin pathway. CEMP1-p4 stimulation upregulated the expression of β-catenin and glycogen synthase kinase 3 beta (GSK-3B) and activated the transcription factors TCF1/7 and Lymphoid Enhancer binding Factor 1 (LEF1) at the mRNA and protein levels. We found translocation of β-catenin to the nucleus in CEMP1-p4-treated cultures. The peptide also penetrates the cell membrane and aggregates around the cell nucleus. Analysis of CEMP1-p4 secondary structure revealed that it has a random-coiled structure. Its biological activities included the induction to nucleate hydroxyapatite crystals. In CEMP1-p4-treated HOMSCs, ALP activity and calcium deposits increased. Expression of Osterix (OSX), Runt-related transcription factor 2 (RUNX2), Integrin binding sialoproptein (IBSP) and osteocalcin (OCN) were upregulated. Altogether, these data show that CEMP1-p4 plays a direct role in the differentiation of HOMSCs to a “mineralizing-like” phenotype by activating the β-catenin signaling cascade.

Isolation and characterization of apical papilla cells from root end of human third molar and their differentiation into cementoblast cells: an in vitro study

“…The differentiation of osteoprogenitors is under the regulation of various signaling pathways, a number of transcriptional factors that fundamentally determine the fate of stem cells toward a “mineralizing-like” phenotype [ 56 , 57 ]. In this study, CEMP1, and CAP, as well as bone-related genes ALP, OCN, DSPP were significantly upregulated [ 58 ].…”

Characterisation and Expression of Osteogenic and Periodontal Markers of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) from Diabetic Knee Joints

“…Reduced cementum thickness was noted in extracted teeth of D vs. ND subjects [ 56 ]. With reports of CEMP-1 (or its derived peptides) inducing osteogenic differentiation of gingival fibroblasts [ 57 ], oral mucosal stem cells [ 58 ] and PDL cells [ 29 ], it seems CEMP-1 functions extend beyond being a marker of cementoblastic differentiation and could be a key molecule for inducing bone and periodontal regeneration both in healthy models and under D conditions.…”

Cementum is key to periodontal tissue regeneration: A review on apatite microstructures for creation of novel cementum‐based dental implants

“…Moreover, a study using the rat fenestration defect model demonstrated that the CEMP1‐p1 peptide induces the deposition of lamellar‐like AEFC, formation of MSC in the presence of cementocytes, and new bone formation near the cementum‐enamel junction (Hoz, López, Zeichner‐David, & Arzate, 2021). In addition to the N‐terminus‐derived peptides of CEMP1, the C‐terminus‐derived peptide of CEMP1 (CEMP‐p4) reportedly promotes mineralization through the Wnt/β‐catenin signaling pathway in human oral mucosa stem cells (Arroyo et al, 2020).…”